Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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1
COMPOUNDS
The present invention relates to novel hydantoin derivatives, processes for
their
preparation, pharmaceutical compositions containing them and their use in
therapy.
Metalloproteinases are a superfamily of proteinases (enzymes) whose numbers in
recent
years have increased dramatically. Based on structural and functional
considerations these
enzymes have been classified into families and subfamilies as described in
N.M. Hooper
(1994) FEBS Letters 354:1-6. Examples of metalloproteinases include the matrix
metalloproteinases (MMPs) such as the collagenases (MMP1, MMP8, MMP13), the
gelatinases (MMP2, MMP9), the stromelysins (MMP3, MMP10, MMP11), matrilysin
(MMP7), metalloelastase (MMP12), enamelysin (MMP19), the MT-MMPs (MMP14,
MMP 15, MMP 16, MMP 17); the reprolysin or adamalysin or MDC family which
includes
the secretases and sheddases such as TNF converting enzymes (ADAM10 and TACE);
the
astacin family which include enzymes such as procollagen processing proteinase
(PCP);
and other metalloproteinases such as aggrecanase, the endothelin converting
enzyme
family and the angiotensin converting enzyme family.
Metalloproteinases are believed to be important in a plethora of physiological
disease
processes that involve tissue remodelling such as.embryonic development, bone
formati6n
and uterine remodelli.n.g during menstruation. This is based on the abili-ty
of the
metalloproteinases to cleave a broad range of matrix substrates such as
collagen,
proteoglycan and fibronectin. Metalloproteinases are also believed to be
important in the
processing, or secretion, of biological important cell mediators, such as
tumour necrosis
factor (TNF); and the post translational proteolysis processing, or shedding,
of biologically
important membrane proteins, such as the low affinity IgE receptor CD23 (for a
more
complete list see N. M. Hooper et al., (1997) Biochem. J. 321:265-279).
Metalloproteinases have been associated with many diseases or conditions.
Inhibition of
the activity of one or more metalloproteinases may well be of benefitin these
diseas-es oy_
conditions, for example: various inflammatory and allergic diseases such as,
inflammation
of the joint (especially rheumatoid arthritis, osteoarthritis and
gout),,inflammation of the
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2
gastro-intestinal tract (especially inflammatory bowel disease, ulcerative
colitis and
gastritis), inflammation of the skin (especially psoriasis, eczema,
dermatitis); in tumour
metastasis or invasion; in disease associated with uncontTolled degradation of
the
extracellular matrix such as. osteoarthritis; in bone resorptive disease (such
as osteoporosis
and Paget's disease); in diseases associated with aberrant angiogenesis; the
enhanced
collagen reinodelling associated with diabetes, periodontal disease (such as
gingivitis),
corneal ulceration, ulceration of the skin, post-operative conditions (such as
colonic
anastomosis) and dermal wound healing; demyelinating diseases of the central
and
peripheral nervous systems (such as multiple sclerosis); Alzlieimer's disease;
extracellular
matrix remodelling observed in cardiovascular diseases such as restenosis and
atheroscelerosis; asthma; rhiuzitis; and chronic obstructive pulmonary
diseases (COPD).
MMP 12, also known as macrophage elastase or nietalloelastase, was initially
cloned in the
mouse by Shapiro et al [1992, Journal of Biological Chemistry 267: 4664] and
in man by
the same group in 1995. MMP12 is preferentially expressed in activated
macrophages, and
has been shown to be secreted from alveolar macrophages from smokers [Shapiro
et al,
1993, Journal of Biological Cheinistiy, 268: 23824] as well as in foam cells
in
atherosclerotic lesions [Matsuinoto et al, 1998, Ain. J. Pathol. 153: 109]. A
mouse model
of COPD is based on challenge of mice with cigarette smolce for six months,
two cigarettes
a day six days a week. Wild-type mice developed pulmonary emphysema afl:er
this
treatment. When MMP12 knock-out mice were tested in this model they developed
no
significant emphysema, strongly indicating that MMP12 is a key enzyme in the
COPD
pathogenesis. The role of MMPs such as MMP12 in COPD (emphysema and
bronchitis) is
discussed in Anderson and Shinagawa, 1999, Current Opinion in Anti-
inflammatory and
Immunomodulatory Investigational Drugs 1 1: 29-38. It was recently discovered
that
smoking increases macrophage infiltration and macrophage-derived MMP- 12
expression
in human carotid artery plaques Kangavari [Matetzky S, Fishbein MC et al.,
Circulation
102: 18 36-39 Suppl. S, Oct 31, 2000].
MMP9 (Gelatinase B, 92kDa Ty_peIV_Colla~enase; 92kDa Gelatinase) is a secreted
protein
_
which was first purified, then cloned and sequenced, in 1989 [S.M. Wilhelm et
al (1989)
J. Biol. Chem. 264 29 : 17213-17221; published erratum in J. Biol. Chem.
(1990) 265
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36 : 22570]. A recent review of MMP9 provides an excellent source for detailed
information and references on this protease: T.H. Vu & Z. Werb (1998) (In:
Matrix
Metalloproteinases, 1998, edited by W.C. Parks & R.P. Mecham, pp. 115 - 148,
Academic Press. ISBN 0-12-545090-7). The following points are drawn from that
review
by T.H. Vu & Z. Werb (1998).
The expression of MMP9 is restricted normally to a few cell types, including
trophoblasts,
osteoclasts, neutrophils and macrophages. However, the expression can be
induced in
these same cells and in other cell types by several mediators, including
exposure of the
cells to growth factors or cytokines. These are the same mediators often
implicated in
initiating an inflammatory response. As with other secreted MMPs, MMP9 is
released as
an inactive Pro-enzyme which is subsequently cleaved to form the enzymatically
active
enzyme. The proteases required for this activation in vivo are not known. The
balance of
active MMP9 versus inactive enzyme is furth.er regulated in vivo by
interaction with
TIMP-1 (Tissue Inhibitor of Metalloproteinases -1), a naturally-occurring
protein. TIMP- 1
binds to the C=tenninal region of MMP9, leading to inhibition of the catalytic
domain of
MMP9. The balance of induced expression of ProMMP9, cleavage of Pro- to active
MMP9
and the presence of TIMP-1 combine to determine the amount of catalytically
active
MMP9 wliich is present at a local site. Proteolytically active MMP9 attacks
substrates
which include gelatin, elastin, and native Type IV and Type V collagens; it
has no activity
against native Type I collagen, proteoglycans or laminins.
There has been a growing body of data implicating roles for MMP9 in various
physiological and pathological processes. Physiological roles include the
invasion of
embryonic trophoblasts through the uterine epithelium in the early stages of
embryonic
implantation; some role in the growth and development of bones; and migration
of
inflammatory cells from the vasculature into tissues.
MMP9 release, measured using enzyme immunoassay, was significantly enhanced in
fluids
and in AM supernantants from untreated asthmatics compared with those from
other
populations [Am. J. Resp. Cell & Mol. Biol., Nov 1997, 17 (5):583-591]. Also,
increased
MMP9 expression has been observed in certain other pathological conditions,
thereby
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implicating MMP9 in disease processes such as COPD, arthritis, tumour
metastasis,
Alzheimer's disease, multiple sclerosis, and plaque rupture in atherosclerosis
leading to
acute coronary conditions such as myocardial infarction.
A number of metalloproteinase inhibitors are known (see, for example, the
reviews of
MMP inhibitors by Beckett R.P. and Whittalcer M., 1998, Exp. Opin. Ther.
Patents,
8 3:259-282; and by Whittaker M. et al, 1999, Chemical Reviews 99(9):2735-
2776).
WO 02/074767 discloses hydantoin derivatives of formula
s R4 Y1
R
R2
R5 A z m kNH
X
y2
that are useful as MMP inhibitors, particularly as potent MMP12 inhibitors.
We now disclose a further group of hydantoin derivatives that are inhibitors
of
metalloproteinases and are of particular interest in inhi.biting MMPs such as
MMP12 and
MMP9. The compounds of the present invention have beneficial potency,
selectivity
and/or pharmacolcinetic properties. The compounds of the present invention are
withi.n the
generic scope of WO 02/074767 but are of a type not specifically exemplified
therein.
In accordance with the present invention, there are provided compounds of
fonnula (I)
R' A'
A 0
~ ~ - N\S. --NH -
0 0 HN4 0
(I)
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wherein
Rl represents H, halogen, CF3 or CH2CN;
5
R2 represents C 1 to 3 allcyl; and
A, Al and B eacli independently represent CH or N;
and pharmaceutically acceptable salts thereof
The compounds of formula (1) may exist in enantiomeric forms. It is to be
understood that
all enantiomers, diastereomers, racemates and mixtures thereof are included
witliin the
scope of the invention.
Compounds of formula (I) may also exist in various tautomeric forms. All
possible
tautomeric forms and mixtures thereof are included within the scope of the
invention.
In one embodiment, Ri represents chloro.
In one embodiment, Rl represents CF3.
In one embodiment, R2 represents methyl or ethyl. In one embodiment, R2
represents
methyl.
In one embodiment, A and Al each represent N. In another embodiinent, A
represents N
and Al represents CH. In another embodiment, A and Al each represent CH.
In one embodiment, B represents N. In another embodiment, B represents CH.
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In one embodiment, Rl represents CF3; R2 represents methyl or ethyl; A and Al
each
represent N; and B represents CH.
s In one embodiment, Rt represents CF3; R2 represents methyl or ethyl; A and
Al each
represent N; and B represents N.
In one embodiment, R1 represents chloro; R2 represents methyl or ethyl; A
represents N
and A1 represents CH; and B represents N.
In one einbodiment, Ri represents chloro; R2 represents methyl or ethyl; and
A, A1 and B
each represent CH.
Unless otherwise indicated, the term "C 1 to 3 allcyP" referred to herein
denotes a straight or
is branched chain alkyl group having from 1 to 3 carbon atoms.. Examples of
such groups
include methyl, ethyl, n-propyl and i-propyl.
Unless otherwise indicated, the term "halogen" referred to herein denotes
fluoro, chloro,
bromo and iodo.
Examples of compounds of the invention include:
(5S)-5-methyl-5-( { [6-[2-(trifluoromethyl)pyrimidin-5-yl]-3,4-
dihydroisoquinolin-2(1H)-
yl]sulfonyl }methyl)imidazolidine-2,4-dione;
(5,S)-5-({ [6-(4-chlorophenyl)-3,4-dihydroisoquinolin-2(1.H)-
yl]sulfonyl}methyl)-5-
methylimidazolidine-2,4-dione;
{4-[2-( { [(4S)-4-methyl-2,5-dioxoimidazolidin-4-yl]methyl} sulfonyl)-1,2,3,4-
tetrahydroisoquinolin-6-yl]phenyl}acetonitrile;
(5S')-5-methyl-5- { [(6-pyridin-3-yl-3,4-dihydroisoquinolin-2(1fl)-
-- --- -- ---
yl~sulfony-I]methyl~iriiidazolidine-2,4-dione;
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(5S)-5-( { [6-(4-chlorophenyl)-3,4-dihydro-2,7-naphthyridin-2( lH)-yl]sulfonyl
}methyl)-5-
methylimidazolidine-2,4-dione;
and pharmaceutically acceptable salts thereof.
Each exemplified compound represents a particular and independent aspect of
the
invention.
The compounds of formula (I) may exist in enantiomeric forms. Therefore, all
enantiomers,
diastereomers, racemates and mixtures thereof are included within the scope of
the invention.
The various optical isomers may be isolated by separation of a racemic
inixture of the
compounds using conventional techniques, for example, fractional
crystallisation, or HPLC.
Alternatively the optical isomers may be obtained by asymmetric synthesis, or
by synthesis
from optically active starting materials.
Where optically isomers exist in the compounds of the invention, we disclose
all individual
optically active forms and combinations of these as individual specific
embodiments of the
invention, as well as their corresponding racemates.
Preferably the compounds of formula (I) have (5S)-stereochemistry as shown
below:
A~ p
R'
K B ~ N~ ~ =,,,, NH
0 0 N~
H 0
Where tautomers exist in the compounds of the invention, we disclose all
individual
tautomeric forms and combinations of these as individual specific embodiments
of the
invention.
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The present invention includes compounds of formula (I) in the form of salts.
Suitable salts
include those formed with organic or inorganic acids or organic or inorganic
bases. Such
salts will normally be pharmaceutically acceptable salts although non-
pharmaceutically
acceptable salts may be of utility in the preparation and purification of
particular
compounds. Such salts include acid addition salts such as hydrocliloride,
hydrobromide,
citrate, tosylate and maleate salts and salts formed with phosphoric acid or
sulphuric acid.
In another aspect suitable salts are base salts such as an alkali metal salt,
for example,
sodium or potassium, an alkaline earth metal salt, for example, calcium or
magnesium, or
an organic amine salt, for example, triethylamine.
Salts of compounds of formula (I) may be formed by reacting the free base or
anotlier salt
thereof with one or more equivalents of an appropriate acid or base.
The compounds of fonnula (I) are useful because they possess phaYmacological
acivity in
animals and are thus potentially useful as pharmaceuticals. In particular, the
compounds of
the invention are metalloproteinase inhibitors and may, thus be used ir1 the
treatment of
diseases or conditions mediated by MMP 12 and/or MMP9 such as asthina,
rhinitis, chronic
obstructive pulmonary diseases (COPD), arthritis (such as rheumatoid arthritis
and
osteoarthritis), atherosclerosis and restenosis, cancer, invasion and
metastasis, diseases
involving tissue destruction, loosening of hip joint replacements, periodontal
disease,
fibrotic disease, infarction and heart disease, liver and renal fibrosis,
endometriosis,
diseases related to the weakening of the extracellular matrix, heart failure,
aortic
aneurysms, CNS related diseases such as Alzheimer's disease and multiple
sclerosis (MS),
and haematological disorders.
In general, the compounds of the present invention are potent inhibitors of
MMP9 and
MMP12. The compounds of the present invention also show good selectivity with
respect
to a relative lac-k of inhibition of various other MMPs such as MMP14.
Accordingly, the present invention provides a compound of foimula_(I),or a_
pharmaceutically acceptable salt thereof, as hereinbefore defined for use in
therapy.
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In another aspect, the invention provides the use of a compound of formula
(I), or a
pharmaceutically acceptable salt thereof, as hereinbefore defined in the
manufacture of a
medicament for use in therapy.
In another aspect, the invention provides the use of a compound of formula
(I), or a
pharmaceutically acceptable salt thereof, as hereinbefore defined in the
manufacture of a
medicament for use in the treatinent of diseases or conditions in which
inhibition of
MMP12 and/or MMP9 is beneficial.
In another aspect, the invention provides the use of a compound of formula
(I), or a
pharmaceutically acceptable salt tliereof, as hereinbefore defined in the
manufacture of a
medicament for use in the treatment of inflammatory disease.
In another aspect, the invention provides the use of a coinpound of formula
(I), or a
pharmaceutically acceptable salt thereof, as hereinbefore defined in the
manufacture of a
medicament for use iri the 'treatment of an obstructive airways disease such
as asthma or
COPD.
In another aspect,, the invention provides the use of a compound of formula
(I), or a
pharmaceutically acceptable salt thereof, as hereinbefore defined in the
manufacture of a
medicament for use in the tireatment of rheumatoid arthritis, osteoartluitis,
atherosclerosis,
cancer or multiple sclerosis.
In the context of the present specification, the term "therapy" also includes
"prophylaxis"
unless there are specific indications to the contrary. The terms "therapeutic"
and
"therapeutically" should be construed accordingly.
Prophylaxis is expected to be particularly relevant to the treat_m.ent of
persons who have
suffered a previous episode of, or are otherwise considered to be at increased
risk of, the
disease or condition in guestion_ Persons at risk of developi__g_ad?articular
disease or
condition generally iuiclude those having a family history of the disease or
condition, or
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those who have been identified by genetic testing or screening to be
particularly
susceptible to developing the disease or condition.
The invention fin=ther provides a method of treating a disease or condition in
which
5 inhibition of MMP12 and/or MMP9 is beneficial which comprises administering
to a
patient a therapeutically effective amount of a compound of formula (1) or a
pharmaceutically acceptable salt thereof as hereinbefore defined.
The invention also provides a method of treating an obstructive airways
disease, for
10 example, asthma or COPD, which comprises administering to a patient a
therapeutically
effective amount of a compound of formula (I) or a pharmaceutically acceptable
salt
thereof as hereinbefore defined.
For the above-mentioned therapeutic uses the dosage administered will, of
course, vary
with the compound employed, the mode of administration, the treatment desired
and the
disorder to be treated. The daily dosage of the compound of formula (1)/salt
(active
ingredient) may be in the range from 0.001 mg/lcg to 75 mg/lcg, in particular
from 0.5
mg/kg to 30 mg/kg. This daily dose may be given in divided doses as necessary.
Typically unit dosage forms will contain about 1 mg to 500 mg of a. compound
of this
invention.
The compounds of formula (I) and pharmaceutically acceptable salts thereof may
be used
on their own but will generally be administered in the form of a
pharmaceutical
composition in which the formula (I) compound/salt (active ingredient) is in
association
with a pharmaceutically acceptable adjuvant, diluent or carrier. Depending on
the mode of
administration, the pharmaceutical composition will preferably comprise from
0.05 to 99
%w (per cent by weight), more preferably fiom 0.10 to 70 %w, of active
ingredient, and,
from 1 to 99.95 %w, more preferably from 30 to 99.90 %w, of a pharmaceutically
acceptable adjuvant, diluent or carrier, all percentages by weight being based
on total
composition. Conventional procedures for the selection and preparation of
suitable
phaYmaceutical formulations are described in, for example, "Pharmaceuticals -
The Science
of Dosage Form Designs", M. E. Aulton, Churchill Livingstone, 1988.
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11
Thus, the present invention also provides a phannaceutical coinposition
comprising a
compound of formula (I) or a pharmaceutically acceptable salt thereof as
hereinbefore
defined in association with a pharmaceutically acceptable adjuvant, diluent or
carrier.
The invention further provides a process for the preparation of a
pharmaceutical
composition of the invention which comprises mixing a compound of formula (I)
or a
pharmaceutically acceptable salt thereof as hereinbefore defined with a
pharmaceutically
acceptable adjuvant, diluent or carrier.
The pharmaceutical coinpositions of this invention may be administered in a
standard
manner for the disease or condition that it is desired to treat, for example
by oral, topical,
parenteral, buccal, nasal, vaginal' or rectal administration or by inhalation.
For these
purposes the compounds of this invention may be formulated by means known in
the art
into the form of, for example, tablets, capsules, aqueous or oily solutions,
suspensions,
emu.lsibns, creams;'ointments; gels, nasal sprays, suppositories, finely
divided powders or
aerosols for inhalation, and for parenteral use (including intravenous,
intramuscular or
infusion) sterile aqueous or oily solutions or suspensions or sterile
emulsions.
In addition to the con-ipounds of the present invention the pharrnaceutical
composition of
this invention may also contain, or be co-administered (simultaneously or
sequentially)
with, one or more pharmacological agents of value in treating one or more
diseases or
conditions referred to hereinabove such as "Symbicort" (trade mark) product.
The present invention further provides a process for the preparation of a
compound of
formula (I) or a pharmaceutically acceptable salt thereof as defined above
which,
comprises:
a) reaction of a compound of formula (II) .
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12 PCT iSE 2005 / o 0 19 1~
O
R2
L S NH
OO N~
H
(II)
wherein R2 is as defined in formula (1) and L1 represents a leaving group,
with a
compound of formula (111) (or a salt thereof)
~ I
A~
B~ NH
(III)
wherein Rl, A, A and B are as defmed in formula (1); or
b) reaction of a compound of formula (V)
R Z O
O p
B~ 10 ~S NH
LG H --~
0
(V)
wherein R2 and B are as defined in formula (1) and LG is a leaving group; with
a boronic
- acid der-ivative-of-form.ula -WI)---
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13
R~ A~
~
A \ I
B(OR)Z
(XII)
wherein Rl, A and A1 are as defined in formula (7); or
c) reaction of a compoun&of formula (IX)
A
I I
.A
B I N 0
0 0 R
(IX)
wherein Rl, R2, A, A1 and B are as defined in formula (1); with ammonium
carbonate and
potassium cyanide;
and optionally thereafter forming a phannaceutically acceptable salt thereof.
In the above process (a), suitable leaving groups Li include halo,
particularly chloro or
trifluoromethylsulfonate. The reaction is preferably performed in a suitable
solvent
optionally in the presence of an added base for a suitable period. of time,
typically 0.5 to 16
h, at ambient to reflux temperature. Typically solvents such as N,N-
dimethylformamide,
pyridine, tetrahydrofu.ran, acetonitrile, N-methylpyrrolidine or
dichloromethane are used.
When used, the added base may be an organic base such as triethylamine,
N,N-diisopropylethylamine, N-methylmorpholine or pyridine, or an inorganic
base such as
an alkali metal carbonate. The reaction is typically conducted at ambient
temperature for
0.5 to 16 h, or until completion of the reaction has been achieved, as
determined by
-------
- chromatographic or spectroscopic methods. Reactions of sulfonyl halides with
varioub
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14
primary and secondary amines are well lcnown in the literature, and the
variations of the
conditions will be evident for those slrilled in the art.
Sulfonylchlorides of formula (II) wherein L1 represents chloro and R2
represents Me are
disclosed in WO 02/074767 and references cited therein. Corresponding
compounds
wherein R2 represents C1 to 3 alkyl may be prepared using analogous methods.
Suitable processes for the preparation of compounds of formula (I) are
described in a
retrosyntlietic way in Scheme 1.
Scheme 1
R\,pI R~pi
p ~ / I p ~ I (RO)ze~~~'~
B\ N%\' 1 O ~ g\ I N~S/ g~ ' N~PO
0 0 R2 //\\
(IXa-c) M-0 o O
(VIIIa-c)
R*pI R' 'pl Rpr
g~ N~S/,r \~ ~ B~ I NH ~ g\ I N~etl
(1) a/\aR' H
(Illa-c) (IVno)
V ~.
LG / I o ~ LG / LG
//\\ lta
O O H
(Va-c)
(Vlflc) (VIIa-c)
0
R~ p~~S/ Y p' Y
o\ORH~'. p~ LG \ B(GR)2
4t) (X[) 01)
In Scheme 1, protecting groups (PG) can be either carbamates (e.g. tert-
butoxycarbamate),
amides (e.g. trifluoroacetyl) or allVl (e.g. tert-butyl or benzyl). Leaving
groups (LG) can
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be either chloride, bromide, iodide or trifluoromethylsulfonate. In the
palladium-catalysed
Suzuki couplings, either boronic acids or pinacolboronates may be used.
Intermediate
(IVa-c) can be prepared by standard Suzuki coupling (Chenz. Rev. 1995, 95,
2457) between
an electrophile (VIIa-c) and a boron reagent (Xll), or the other way around,
between an
5 electrophile (XI) and a boron reagent (VIIla-c). The latter can be obtained
from (VIIa-c)
using standard Miyaura conditions (J. Org. Chefn.1995, 60, 7508-7510).
Deprotection of
(1Va-c) either by hydrogen chloride in metlianol (PG = tert-butoxycarbonyl) or
refluxing 1-
chloroethyl chloroformate/ refluxing methanol (PG = ter=t-butyl or berizzyl)
(Synlett. 1993,
195-196) gives amine (IIIa-c) as a hydrochloride salt. The free base can be
obtained by
10 treatinent of (Illa-c) with base and extraction with an organic solvent
such as ethyl acetate
or toluene. Reacting (Illa-c) either as a salt or base in a suitable solvent
(e.g. acetonitrile,
tetrahydrofuran, N-methylpyrrolidine or NN-dimethylfoilnamide) with the
sulfonyl
chloride (II) in the presence of a tertiary amine (e.g. triethylamine,
pyridine or
N,N-diisopropylethylamine) for 0.5 to 16 hours produces compounds of formula
(I).
An alternative route to compounds of formula (I) from intermediate (Illa-c)
via
methanesulfonamide (Xa-c) and ketone (IXa-c) has been previously described (WO
02/074767). Briefly, treatment of (IIIa-c) witli, methansulfonyl chloride and
a tertiary
amine (e.g. triethylamine, pyridine or.N,N-diisopropylethylamine) in a
suitable solvent
(e.g. dichloromethane or tetrahydroffaran) produces the methansulfonamide (Xa-
c) which
in turn can be transformed into the ketone (IXa-c) using standard procedures.
Heating
ketone (IXa-c) with ammonium carbonate and potassiuin cyanide in 50% aqueous
ethanol
in a sealed vial at 80-90 C for 1 to 5 hours gives a racemic hydantoin that
can be resolved
by chiral chromatography (e.g. on OD-H with 100% ethanol).
In a third route, intermediate (VIIa-c) is deprotected as described above to
give amine
(VIa-c) as a hydrochloride salt. The free base can be isolated by treatment
with base and
extraction with an organic solvent e.g. etliyl acetate or toluene. Reacting
(VIa-c) either as a
salt or base in a suitable solvent (e.g. acetonitrile, tetrahydrofuran, N-
methylpyrrolidine or
N,N-dimethylformamide) with sulfonyl chloride (lI) in the presence of a
tertiarv amine
(e.g. triethylamine, pyridine or N,N-diisopropylethylamine) for 0.5 to 16
hours produces
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16
chiral sulfonamide (Va-c). The latter can be coupled with boron reagent (XII)
using
standard Suzuld conditions to give compounds of foi7nula (I).
Intermediates (VIIa-b) are conveniently prepared using the following methods.
The 1,2,3,4-tetrahydroisoquinoline intermediate (Vlla)
Methods for the synth.esis of 1,2,3,4-tetrahydroisoquinolines are well lcnown
in the
literature. The classical route is the Pomeranz-Fritz reaction of
benzaldehydes with a
diacetal protected aminoacetaldehyde (Org. React. 1951, 6, 191) yielding the
isoquinoline
nucleus which upon catalytical reduction gives 1,2,3,4-tetrahydro-
isoquinolines. Another
route is the Bischler-Napieralsld reaction (Org. React. 1951, 6, 74) of a
carbamate of
2-phenylethanamines with phosphoryl chloride in refluxing toluene or xylenes.
Reduction
of the resulting cyclic benzamide with lithiuin aluminium hydride in
tetrahydrofuran (1.
Med. Chem. 1987, 30(12), 2208-2216) or diborane in tetrahydrofuran (J. Med.
Chenz.
1980, 23(5), 506-511) affords the 1,2,3,4-tetrahydroisoquinoline. A variation
of the
Bischler-Napieralski reaction is the Pictet-Spengler synthesis (Org. React.
1951, 6, 151). In
this reaction amides, carbamates or sulfonamides of 2-phenylethanaYnines are
heated with
paraformaldehyde and strong proton acids (e.g. trifluoroacetic acid, sulfiu7c
acid) or Lewis
acids in a solvent (e.g. dichloromethane, toluene, formic acid) to give the
1,2,3,4-
tetrahydroisoquinoline in a single step (Tetrahedron 2002, 58(8), 1471-1478).
Scheme 2
Br /~ a, b r Br / Br
/ d Br W"COOBu-t
\ HtTz \ I NICOCF\ I ~~ Route A (+ B-Br-isomer) VIIn 03 and 131 = CH
and LG = Br)
Reagents:
a) (CF3CO)2O, Et3N; +4 C. b) (HCHO),,, H2SO4, HOAc; RT. c) NaBH4, E?OH; RT or
NHj (conc), EIOH, heat
d) (t-BuOCO),O, Et3N, DCNd, RT.
Preferably the 1,2,3,4-tetrahydroisoquinoline intermediate (VIIa) is
synthesised by Route
A shown in Scheme 2. This route is a Friedel-Craft-type reaction of N-[2-(3-
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17
bromophenyl)ethyl]-2,2,2-trifluoroacetamide with formaldehyde and sulfuric
acid in acetic
acid (Tetrahedron Lett. 1996, 3 7(31), 5453-5456) giving a mixture of the 6-
bromo- and
8-bromoisomer in a ratio of 3 to 1. Replacement of the trifluoroacetamide
group with a
BOC-group gives (Vlla). The regioisomers are not conveniently separated at
this stage.
The 1,2,3,4-tetrahydro-2,7-naphthyridine intermediate CVllb)
In contrast to the 1,2,3,4-tetrahydroisoquinolines, there are rather few
examples of
synthetic methods for 1,2,3,4-tetrahydro-2,7-naphthyridines in the literature.
One
importa.nt method to prepare 1,2,3,4-tetrahydro-2,7-naphthyridine is the regio-
selective
catalytic reduction of 2,7-napllthyridine (Eur. J. Med. Chena. Ther. 1996,
31(11), 875-888).
The synthesis of 2,7-naph.thyridine and some derivatives thereof has been
described in the
literature. One classical route involves several steps and starts with the
acid catalysed
condensation of malononitrile with diethyl 1,3-acetonedicarboxylate (J. Chein.
Soc. 1960,
3513-3515; see also J. Het.erocycl. Chern. 1970, 7, 419-421). A slightly
different route to
2,7-naphthyridine involves oxidation of 4-formyl-2,7-naphth.yridine to give
2,7-naphthyridine-4-carboxylic acid followed by decarboxylation (Syntlzesis
1973, 46-47).
A completely different method is based on the internal Diels-Alder reaction of
N-(ethoxycarbonyl)-N-(but-3-ynyl)amino-methylpyrazine and gives a mixture of
1,2,3,4-
tetrahydro-2,7-naphthyridine and 5,6,7,8-tetrahydro-1,7-naphthyridine after
hydrolysis of
the carbamate group (WO 02/064574).
Scheme 3
Route B
SiMe~
Me0 / a, b Meo /II e MeO d Me0
N,, 0 Nz i 0 i0 N, A, N
Teo h xo , g xo ~ f Meo
N\ NIN~ I N. N~ NH N, NH
COOBu-t COOBu-[
xHBr
Reagents~--
a) LiCH}NCH2CH,N(CH}; Z, THF, -70 C, b) n-BuLi in he:canes, -70 C, then I,. c)
TMS-acetylone, PdCI,(PPh3)2, CuI, Et3N, THF, 60 C.
d) 7 M NH3, EtOH, 80 C. o)Hõ PtOõ HOAc. f) 48"/u HBr (aq), 120 C. g) (BOC)2O1
Et}N, H,O, THF. h) TEi0, PhMe, 30% P3P04.
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18
Preferably the 1,2,3,4-tetrahydro-2,7-naphthyridine inteimediate (Vllb) can be
synthesised
as shown in Schemes 3 and 4. In Route B, commercially available
6-methoxynicotinaldehyde is treated successively with the lithium salt of
NN,N'-trimethylethylenediamine, then n-BuLi in hexanes and finally iodine to
afford the
4-iodo-6-methoxynicotinaldehyde (cf. Tetrahedron Lett. 1993, 34(39), 6173-
6176). The
iodo compound is coupled with trimethylsilylacetylene under usual Sonagashira-
Hagihara
conditions (Synthesis 1980, 627-630) al.ld the resulting 6-methoxy-4-
[(trimethylsilyl)ethynyl]nicotinaldehyde is condensed with ammonium hydroxide
in
ethanol to give 3-methoxy-2,7-naphthyridine (Synthesis 1999, 2, 306-311).
Regioselective
catalytical reduction (cf. Eur. J. Med. Chern. Ther. 1996, 31(11), 875-888)
affords
.6-methoxy-1,2,3,4-tetrahydro-2,7-naphthyridine. Demethylation and N-
protection with
BOC-anh.ydride and finally treatment of the resulting tert-butyl 6-hydroxy-3,4-
dihydro-
2,7-naphthyridine-2(1H)-carboxylate with triflic anhydride in a two-phase
system gives
(Vllb).
Scheme 4
Roure C
Me0 / Me0 b Ma0 c Me0 / ,0 a
N~ I ~O - - N\ I N'
Bu-t i IIu-i
d
Tf0 f HO e Me0
~- E
~ ~ I rl.
Bu-t Bu-[ Bu-t
Reagents:
a) n-BuLi, THF, -70 C then DMF, -70~C to RT. b) t-BuNHz, DCM, 3A mol. sieves.
e) Li-TMP, -200C then DMF, -20 to -10"C.
d) Ns13HrCN, MeOH, HOAc; RT. e) 48% HBr (aq), re[lux; work-up with KZCO3 (sq).
f) TfzO, pyridinc +4 C.
In Route C, commercially available 5-bromo-2-methoxy-4-methylpyridine in
anliydrous
_ tetrahydrofuran is metallated with n_BuLia.nd thentreatedwithN, N-
climethylformauaide to
afford 6-methoxy-4-methylnicotinaldehyde. This was converted to the tert-
butylimine with
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19
tert-butylamine in dichloromethane. Metallation with lithiuin 2,2,6,6-
tetramethylpiperidide
(Li-TMP) (cf. J. Org. Che a.1993, 58, 2463-2467) and addition of
N,N-dimethylformamide affords the iminoacetaldehyde which is reduced with
sodium
cyanoborohydride in methan.ol to give 2-tert-butyl-6-methoxy-1,2,3,4-
tetrahydro-2,7-
naphthyridine. Cleavage of the methyl group witli refluxing 48% hydrobromic
acid and
treatment with triflic anhydride in the presence of base gives (VIIb)
protected as the
tert-butylamine.
It will be appreciated by those skilled in the art that in the processes of
the present
invention certain potentially reactive functional groups such as hydroxyl or
amino groups
in the starting reagents or intennediate compounds may need to be protected by
suitable
protecting groups. Thus, the preparation of the compounds of the invention may
involve, at
various stages, the addition and removal of one or more protecting groups.
Suitable protecting groups and details of processes for adding and removing
such groups
are described in'Protective Groups in Organic Chemistry', edited by J.W.F.
McOmie,
Plenum Press (1973) and'Protective Groups in Organic Synthesis', 3rd edition,
T.W.
Greene and P.G.M. Wuts, Wiley-Interscience (1999).
The compounds of the invention and intermediates thereto may be isolated from
their
reaction mixtures and, if necessary further purified, by using standard
techniques.
The present invention will now be further explained by reference to the
following
illustrative examples.
General Methods
'H NMR and 13C NMR spectra were recorded on a Varian Inova 400 MHz or a Varian
Altercury-VX 300 MHz inst_riament. The central peaks of chloroform-d (6H 7.27
ppm),
dimethylsulfoxide-d6 (SH 2.50 ppm), acetonitrile-d3 (SH 1.95 ppm) or methanol-
d4 (SH 3.31
_ 30 __ _ppm) were used_as
intemal.references_Column_chromatography_was._carried_out using_
silica gel (0.040-0.063 mm, Merck) with a slight over-pressure (0.2-0.4 bars)
applied on
the column. A Kromasil KR-100-5-C1$ column (250 x 20 mm, Akzo Nobel) and
mixtures
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of acetonitrile/water with 0.1 % TFA at a flow rate of 10 mL/min were used for
preparative
HPLC. Unless stated otherwise, starting materials were commercially available.
All
solvents and commercial reagents were of laboratory grade and were used as
received. The
organic phases from extractions were dried over anhydrous sodium sulfate if
not stated
5 otherwise. Organic phases or solutions were concentrated by rotary
evaporation. Yields
were not optimised.
The following method was used for LC-MS anal sis:
Instrument Agilent 1100; Column Waters Synzrnetry 2.1 x 30 mm; Mass APCI; Flow
rate
io 0.7 mL/min; Wavelength 254 or 220 nm; Solvent A: water + 0.1% TFA; Solvent
B:
acetonitrile + 0.1% TFA ; Gradient 15-95%/B 2.7 min, 95% B 0.3
The following method was used for GC-MS analysis:
Instrument Hewlett Paclcard 5890 Series II; Column Agilent HP-5 (30 in x 0.32
mm ID);
15 Mass selective detector Hewlett Packard 5971 Series ; Pressure 55 kPa He;
Oven program
100 C (3 min) to 300 C, 25 C/
Abbreviations:
20 BOC-anhydride di-tert-butyl dicarbonate
n-BuLi n-butyl lithium
DCM dichloromethane
DIPEA N,N-diisopropylethylamine
DMF N,N-dimethylformamide
DMSO dimethylsulfoxide
EtOAc ethyl acetate
EtOH ethanol
GC-MS gas chromatography- mass spectrometry -
LDA lithium diisopropylarnide
MeOH methanol LC-MS liquid chromatography- mass spectroscopy
PdCl2 x dppf 1,1'-bis(diphenylphosphino)ferrocene palladium(II)dichloride
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21
RT room temperature, normally 20 to 22 C
TEA triethylamine
THF tetrahydrofuran
TBME tert-butyl methyl ether
TFA trifluoroacetic acid
Triflic anhydride trifluoromethanesulfonic anhydride (Tf20)
Example 1 (5S)-5-Methyl-5-({[6-[2-(trifluoromethyl)pyrimidin-5-yl]-3,4-
dih droisoquinolin-2(1H)-yl]sulfonyl}methyl)imidazolidine-2,4-dione
F
F
F ~ ~N
N~
H
N1~0 N
O H O
[(4S)-4-Methyl-2,5-dioxoimidazolidin-4-yl]methanesulfonyl chloride (0.0295 g,
0.13
mmol) in dry THF (0.60 mL) was added dropwise to a stirTed solution of
6-[2-(trifluoromethyl)pyrimidin-5-yl]-1,2,3,4-tetrahydroisoquinoline (0.039 g,
0.14 mmol),
DIPEA (0.034 mL, 0.20 mmol) and dry THF (0.60 mL) at ice-bath temperature.
After the
addition was complete the solution was stirred at RT for 2 h and then taken up
in water-
brine and extracted twice with EtOAc. The combined organic phases were washed
with
brine, dried, filtered and concentrated to give a crude product. Purification
by preparative
HPLC afforded 0.050 g (76%) of the title compound as a white solid.
LC-MS m/z 470 (M+l);
1H NMR (CD3CN) S 9.19 (s, 214), 8.51 (br s, 1H), 7.62 (s, 1H), 7.61 (dd, 1H),
7.36 (d,
1H), 6.33 (br s, 114), 4.51 (s, 2H), 3.57 (t, 2H), 3.52 (d, 1H), 3.42 (d, 1H),
3.04 (t, 2H) and
1.48 (s, 3H) ppm.
The starting materials were prepared as follows:
6-[2-(Trifluorometh~)pyrimidin-5-yl]-1,2,3,4-tetrahydroisoquinoline
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22
tert-Buty16-[2-(trifluorom ethyl)pyrimidin-5-yl]-3,4-dihydroiso quinoline-2
(1H)-
carboxylate (0.051 g, 0.13 mmol) was stirred in TFA (1.0 mL) and DCM (1.0 mL)
at RT
overnight, then concentrated twice, the second time with added toluene (5 mL),
to afford
the trifluoroacetate salt.
LC-MS rizlz 280 (M+1);
1H NMR (CD3CN) S 9.25 (s, 2H), 7.73 (m, 2H), 7.44 (d, 1H), 4.45 (s, 2H), 3.56
(t, 2H)
and 3.24 (t, 2H) ppm.
The crude product was taken up in 1M sodium carbonate solution (10 mL) and
extracted
twice with EtOAc. The combined organic phases were washed witli brine, dried,
filtered
and concentrated to give 0.039 g (100%) of the title product as a white solid.
2-(Trifluorometh&I)pyriinidin-5-yl trifluoromethanesulfonate
Triflic anhydride (13.9 g, 85 nnnol) in dry DCM (70 mL) was added slowly to an
ice-cold
solution of 2-(trifluoromethyl)pyrimidin-5-ol (13.9, 85 mmol) (US 4,558,039),
DIPEA (16
1s mL, 93 mmol) and dry DCM ( 260 mL) at such a rate that the temperature was
kept
between 4 C and 6 C. After the addition was complete, the solution was'stirred
for 2.5 h
at 4 C and then allowed to warm to RT. Water (50 mL) and 1M phosphoric acid
(4.5 mL)
were added and the phases were washed and separated. The organic phase was
washed
successively with water and saturated sodium bicarbonate, dried, filtered and
carefully
concentrated by rotary evaporation (pressure 300-400 mbar). The dark-red oil
was purified
by column chromatography with EtOAc-heptanes (1:8 through 1:4) as eluent to
give 22.5 g
(90%) of the title product as a colourless oil that crystallised in the cold.
Alternatively, the
product could be purified by distillation, b.p. 75-77 C/10 mbar.
H NMR (CDC13) 6 8.90 (s, 2H) ppm.
ter t-Buty16-[2-(trifluoromethXl)p, rimidin-5-yl]-3,4-dihydroisoquinoline-2
1HZ
carbox. late
A 4:1 mixture (0.10 g, 0.28 mmol) of ter=t-butyl6-(4,4,5,5-tetramethyl-1,3,2-
d.ioxaborolan-
2-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate and tert-butyl8-(4,4,5,5-
tetramethyl-
--
so 1~3,2=dioxaborolan=2=y1)=3,4=dihydroisoquinoline=2(11~-carboxylate,
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23
2-(trifluoromethyl)pyrimidin-5-yl trifluoromethanesulfonate (0.083 g, 0.28
mmol), PdC12 x
dppf (0.0048 g), 2M sodium carbonate (1.1 mL), toluene (4.0 mL) and EtOH (1.0
mL) was
purged with dry argon for ten minutes then heated in a sealed vial at 81 C
for 6 h. The
black solution was filtered through glass-wool, taken up in water-brine and
washed twice
with EtOAc. The combined organic phases were dried, filtered and concentrated
with silica
(5 g). Column chromatography with EtOAc-heptanes (1:8 through 1:5) gave 0.051
g (48%)
of the title product as white solid.
LC-MS rrr/z 380 (M+1);
1H NMR (CDC13) 8 9.06 (s, 2H), 7.44 (dd, 1H), 7.38 (br s, 1H), 7.30 (d, 1H),
4.66 (s, 2H),
3.71 (t, 214), 2.95 (t, 214), and 1.51 (s, 9H) ppm.
tert-Buty16-(4 4 5 5-tetramethyl-1 3 2-dioxaborolan-2-yl)-3 4-
dihydroisoquinoline-2(1H)
carbox late
A 3:1 mixture (0.49 g, 1.6 mmol) of teNt-butyl6-bromo-3,4-dihydroisoquinoline-
2(1H)-
1s carboxylate and tert-butyl8-bromo-3,4-dihydroisoquinoline-2(1H)-
carboxylate, bis(pinacolato)diborane (0.45 g, 1.8 mmol), PdC12 x dppf (0.039
g, 0.048
mmol), potassiuin acetate (0.48 g, 4.8 mmol) and DMF (8.0 mL) was heated at 81
C
overnight. The solvent was evaporated, the residue taken up in water-brine and
washed
twice with EtOAc. The organic phase was dried, filtered and concentrated.
Column
chromatography with EtOAc-heptanes (1:10 through 1:4) gave 0.24 g of a 4:1
niixture of
the title product and tert-butyl 8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
yl)-3,4-
dihydroisoquinoline-2( lH)-carboxylate.
1H NMR (CDC13) S 7.62 (d, 1H), 7.60 (s, 1H), 7.13 (d, 1H), 4.59 (s, 2H), 3.64
(t, 2H), 2.85
(t, 2H), 1.50 (s, 9H) and 1.35 (s, 12H) ppm (6-isomer).
1H NMR (CDC13) 8 7.69 (d, 1H), 7.24-7.14 (m, 214), 4.88 (s, 214), 3.64 (t,
2H), 2.85 (t,
214), 1.50 (s, 9H) and 1.35 (s, 12H) ppm (8-isomer).
tert-Butyl 6-bromo-3,4-dihydroisoguinoline-2(1H)-carbox vlate
6-Bromo-2-(trifluoroacetyl) 1,2,3,4-tetrahydroisoquinoline was prepared in two
steps from
[2-(3-bromophenyl)ethyl]amine (4.0 g, 20 mmol) following the procedure of
Stokker
(Tetrahedron Lett. 1996, 3 7(31), 5453-5456). Column chromatography with EtOAc-
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24
heptanes (1:10 through 1:6) gave 2.3 g (7.5 mmol) of a 3:1 mixture of 6-bromo-
2-
(trifluoroacetyl)-1,2,3,4-tetrahydroisoquinoline and 8-bromo-2-(trifluoro-
acetyl)-1,2,3,4-
tetrahydroisoquinoline.
iH NMR (CDC13) S 7.62 (d, 1H), 7.60 (s, 1H), 7.13 (d, 1H), 4.59 (s, 2H), 3.64
(t, 2H), 2.85
(t, 2H) and 1.50 (s, 9H) and 1.35 (s, 12H) ppm (6-isomer).
iH NMR (CDC13) 8 7.69 (d, 1H), 7.24-7.14 (m, 2H), 4.88 (s, 2H), 3.64 (t, 2H),
2.85 (t, 2H)
and 1.50 (s, 9H) and 1.35 (s, 12H) ppm (8-isomer).
The above material was stirred with absolute EtOH (100 mL) and 25% ammonium
hydroxide (10 mL) at 60 C for 4 h. More 25% ammonium hydroxide (15 mL) was
added
and stirring continued at RT overnight. The volatiles were evaporated to leave
the crude
amine as a white solid.
LC-MS m/z 212, 214 (M+l ).
Dry THF (50 mL) and DIPEA (1.3 mL, 7.5 mmol) were added followed by BOC-
anhydride (1.8 g, 8.2 nunol). The mixture was stirred at RT overnight. The
volatiles were
evaporated and the residue was taken up in water. The pH was adjusted to 2
with 1M
phosphoric acid and the product was extracted twice with EtOAc. The conibined
organic
phases were washed with brine made slightly alkaline with saturated sodium
bicarbonate,
dried, filtered and concentrated. The crude product was purified by column
chromatography with EtOAc-heptanes (1:50 through 1:20) to give 2.24 g (96%) of
a 3:1
mixture of the title product and teNt-butyl8-bromo-3,4-dihydroisoquinoline-
2(1H)-
carboxylate.
LC-MS mlz 256, 258 (M-56);
iH NMR (CDC13) S 7.31 (dd, 1H), 7.30 (br s, 1H), 6.98 (d, 1H), 4.52 (s, 2H),
3.63 (t, 2H),
2.81 (t, 2H) and 1.50 (s, 9H) ppm (6-isomer .
1H NMR (CDC13) S 7.42 (dd, 1H), 7.12-7.01 (m, 214), 4.55 (s, 2H), 3.64 (t,
211), 2.84 (t,
2H) and 1.51 (s, 9H) ppm (8-isomer).
Example 2 (5S)-5-({F6-(4-Chlorophenyl)-3,4-dih droisoquinolin-2(1H)-
_
- --------
yl] sulfoUl} methyl)-5 -meth=ylimi dazolidine-2,4-dione
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ci /
o
H
N, i0 N
oS~.u
0 0
H
(5S)-5- { [(6-Bromo-3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl]methyl}-5-methyl-
imidazolidine-2,4-dione (0.016 g, 0.040 mmol), 4-chlorophenylboronic acid
(0.0072 g,
5 0.045 mmol), PdC12 x dppf (0.0030 g), 2M sodium carbonate (0.15 mL), toluene
(0.80 mL)
and EtOH (0.20 mL) were stirred in a sealed vial at 95 C for 17 h. The
solvent was
evaporated and the residue was tal(en up in water. The solution was acidified
with 10%
HOAc to pH 6 and then extracted twice with EtOAc. The combined organic phases
were
washed with brine-saturated sodium bicarbonate, dried, filtered and
concentrated to give a
10 crude product.
LC-MS n2/z 434 (M+1).
Purification by preparative HPLC afforded 0.0080 g (46%) of the title compound
as a
white solid.
1H NMR (CD3CN) S 8.53 (br s, 1H), 7.62 (m, 2H), 7.46 (m, 4H), 7.23 (d, 1H),
6.34 (br s,
15 1H), 4.45 (s, 214), 3.53 (m, 2H), 3.49 (d, 1H), 3.39 (d, 1H), 2.99 (m, 2H)
and 1.46 (s, 3H)
ppm=
The compounds of Examples 3 and 4 were prepared using the general method of
Example
2.
Example 3 14-F2-({r(4S)-4-Methyl-2,5-dioxoimidazolidin-4-yl]methyllsulfonyl)-
1,2,3,4-tetrahydroisoquinolin-6-yllphenLI} acetonitrile
N
N, 1~0 O H
OS....a N 0
11_
H
White solid.
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26
LC-MS nZ/z 439 (M+1);
iH NMR (CD3CN) S 8.61 (br s, 1H), 7.65 (m, 2H), 7.48 (m, 2H), 7.43 (m, 2H),
7.23 (d,
1H), 6.38 (br s, 1H), 4.46 (s, 2H), 3.87 (s, 2H), 3.53 (m, 2H), 3.50 (d, 1H),
3.40 (d, 1H),
3.00 (m, 2H) and 1.46 (s, 3H) ppm.
Example 4 (SS)-S-Methyl-5-{F(6-pyridin-3-yl-3,4-dih droisoquinolin-2(1H)-
yl)sulfonLl]methyl} imidazolidine-2,4-dione
N~ o
i,0 N
u O
O ~
White solid.
LC-MS na/z 401 (M+l);
1H NMR (CD3CN) S 8.98 (br s, 1H), 8.71 (m, 1H), 8.54 (d, 2H), 7.89 (m, 1H),
7.56 (m,
2H), 7.34 (m, 1H), 6.34 (br s, 1H), 4.49 (s, 2H), 3.55 (m, 2H), 3.52 (d, 1H),
3.41 (d, 1H),
3.03 (m, 2H) and 1.47 (s, 3H) pp.in.
The starting material was prepared as follows:
(5S)-5-(f [6-Bromo-3,4-dihydroisoquinolin-2(1ffi-yl]sulfon l}methyl)-5-
methylimidazolidine-2,4-dione
A 3:1 rnixture (0.44 g, 1.4 mmol) of 6-bromo-2-(trifluoroacetyl)-1,2,3,4-
tetrahydroisoquinoline and 8-bromo-2-(trifluoroacetyl)-1,2,3,4-
tetrahydroisoquinoline
(prepared according to Tetrahedron Lett. 1996, 37(31), 5453-5456) was stirred
in ethanol
(10 mL) containing a few drops of 25% ammonium hydroxide at RT. After 2.5 h,
the
solution was concentrated, dissolved in dry THF (1.0 mL) under argon and
cooled on an
ice-bath. DIPEA (0.41 mL, 2.4 mmol) was added followed by a solution of [(4S)-
4-methyl-
2,5-dioxoimidazolidin-4-yl]methanesulfonyl-chloride-(0.-27-g, 4:2 mmol)-and-
dry THF (1.0 -
mL). The mixture was stirred at RT for 1 h and then concentrated. The crude
product was
taken up in water and extracted twice with EtOAc. The combined organic phases
were
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27
washed with brine, dried, filtered and concentrated to give 0.55 g of a
mixture of (5,S)-5-
( { [6-bromo-3,4-dihydroisoquinolin-2(1H)-yl]sulfonyl}methyl)-5-
methylimidazolidine-2,4-
dione and (5S)-5-({[8-bromo-3,4-dihydroisoquinolin-2(lH)-yl]sulfonyl}methyl)-5-
methylimidazolidine-2,4-dione. The regioisomers were separated by preparative
HPLC.
(5S)-5-({[8-Bromo-3,4-dih droisoquinolin-2(1.H)-yl]sulfon 1}methyl)-5-
methylimidazolidine-2,4-dione (eluting first)
Yield: 0.13 g of a white solid.
LC-MS nalz 402/404 (M+1), 419/421 (M+18);
1H NMR (CD3CN) 8 8.48 (br s, 1H), 7.48 (m, 1H), 7.21 (m, 1H), 7.14 (m, 1H),
6.31 (br s,
1H), 4.36 (s, 2H), 3.48 (m, 4H), 2.95 (m, 2H) and 1.46 (s, 314) ppm.
(5S)-5-( f f6-BroYno-3,4-dih droquinolin-2(1H)-yllsulfonyl}meth 1~
methylimidazolidine-2,4-dione (eluting second)
1s Yield: 0.25 g of a white solid.
LC-MS m/z 402/404 (M+1), 419/421 (M+18);
I H NMR (CD3CN) S 8.47 (br s, 1H), 7.38 (m, 1H), 7.36 (m, 1H), 7.08 (m, 1H),
6.29 (br s,
1H), 4.36 (s, 2H), 3.48 (m, 2H), 3.47 (d, 1H), 3.37 (d, 1H), 2.92 (m, 2H) and
1.45 (s, 3H)
ppm.
Example 5 (5,S)-5- {[6-(4-Chlorophenyl)-3,4-dihydro-2,7-naphthyridin-2(1H)_
yl] sulfonyl } methyl)-5 -methylimidazolidine-2, 4-dione
ci /
o
Nz~
N i0 ~
.,.~~ H
O O
[(4,S')-4-Methyl-2,5-dioxoimidazolidin-4-yl]methanesulfonyl chloride (0.086 g,
0.38 mmol)
in-anhydrous NMP-(0:50 -mL-)-was added-dropwise to-a stirred-solution of-
6-(4-chlorophenyl)-1,2,3,4-tetrahydro-2,7-naphthyridine (0.046 g, 0.19 rnmol),
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28
DIPEA (0.066 mL, 0.38 minol) and anhydrous NMP (1.5 mL) at RT. After the
addition
was complete the solution was stirred at RT for 1.5 h, then diluted with water
(1 mL) and
purified by preparative HPLC to afford 0.0070 g (8%) of the title compound as
a white
solid.
LC-MS rn/z 435, 436 (M+1);
iH NMR (DMSO-d6) 8 10.8 (s, 1H), 8.49 (s, 1H), 8.10 (d, 2H), 8.06 (s, 1H),
7.84 (s, 1H),
7.54 (d, 2H), 4.45 (s, 2H), 3.61 (d, 1H), 3.48 (d, 1H), 3.47 (t, 2H), 2.98 (t,
2H) and 1.34 (s,
3H) ppm.
The starting materials were prepared as follows:
6-(4-Chlorophenyl)-1,2,3,4-tetrahydro-2,7-naphthridine
tert-Butyl 6- { [(trifluoromethyl)sulfonyl]oxy} -3,4-dihydro-2,7-naphthyridine-
2(1H)-
carboxylate (0.69 g, 1.8 mmol), 4-chlorophenylboronic acid (0.39 g, 2.5 mmol),
PdC12 x
dppf (0.050 g), saturated sodium carbonate (2 mL), EtOH (4 mL) and toluene (4
mL) were
stirred at 80 C for 6 h. The solution was cooled to RT, talcen up in water
(10 mL) and
extracted with EtOAc (25 mL). The combined organic phases were washed with
brine,
dried, filtered and concen.trated. Purification by column chromatography with
EtOAc-
heptanes (1:1) as eluent gave 0.065 g (10%) of tert-butyl6-(4-chlorophenyl)-
3,4-dihydro-
2,7-naphthyridine-2(1R)-carboxylate.
LC-MS nilz 345 (M+l).
This material was dissolved in MeOH (2 mL) and acetyl chloride (0.2 mL) was
slowly
added. After stirring at 40 C overnight, the solution was concentrated, the
residue was
taken up in 1 M sodium hydroxide (10 mL) and extracted with EtOAc-ether (1:1)
(4 x 30
mL). The combined organic phases were dried, filtered and concentrated to give
0.046 g
(100%) of the crude title compound.
LC-MS mlz 245 (M+1).
tert-Butyl 6-{[(trifluoromethyl)sulfonyl]oxY -3,4-dihydro-2,7-naphthyridine-
2(1H)-
--'30---- -carboxylate-
Crude 3-methoxy-2,7-naphthyridine (prepared from 4.4 mmol of 6-methoxy-4-
[(trimethylsilyl)ethynyl]nicotinaldehyde) was hydrogenated (30 psi pressure)
at RT over
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29
Pt02 (approx. 0.1 g) in HOAc (25 mL) for 2.5 h. The solution was filtered
through a Celite
pad and the clear filtrate was concentrated by freeze-diying to give crude 6-
methoxy-
1,2,3,4-tetrahydro-2,7-naphthyridine as the acetate salt.
LC-MS fnlz 165 (M+1).
This material was refluxed in 48% hydrobromic acid for 10 h. The volatiles
were
evaporated and the residue was dried under vacuum and 45 C to give approx.
0.70 g. of
crude 5,6,7,8-tetrahydro-2,7-naphthyridin-3-ol hydrobromide.
LC-MS rnlz 151 (M+1).
This material (approx. 4.8 mmol) was dissolved in water (13 mL) and treated
with TEF
(33 inL), Et3N (0.85 mL, 6.0 mmol) and BOC-anhydride (1.6 g, 7.3 mmol) at RT.
After
stiiTing at the same temperature for 6 h the solution was concentrated to one
third of its
original volume and the residue was taken up in water and extracted three
times with
EtOAc. The combined organic phases were dried, filtered and concentrated to
give 0.80 g
(67% crude yield) of tert-butyl6-hydroxy-3,4-dihydro-2,7-naphthyridine-2(111)-
i5 carboxylate as a white solid.
LC-MS mlz 251 (M+1), 195 (M-55).
This material (approx. 5.4 mmol) was dissolved in a two-phase system of
toluene (20 mL)
and 30% aqueous tripotassium orthophosphate (20 mL) and treated with triflic
anhydride
(1.6 mL, 6.8 mniol) at 4 C [Org. Lett. 2002, 4(26), 4717-4718]. The ice-bath
was removed
and the stirring was continued for 2 h at RT after which the two phases were
separated.
The aqueous phase was washed once with toluene. The combined organic phases
were
washed with brine, dried and concentrated. Purification by column
chromatography with
EtOAc-heptanes (2:1) as eluent gave 0.45 g (17 % yield) of the title product.
LC-MS rnlz 383 (M+1), 283 (M-99).
3-Methox -~2,7-naphthyridine
To a stirred solution ofN,N,N'-trimethylethylenediamine (1.9 mL, 15 mmol) in
anhydrous
THF (65 mL) under argon at -70 C was slowly added 1.6M n-BuLi in hexanes (9.0
mL,
14 mmol). After stirring at -70 C for 15 minutes, 6-methoxy-nicotinaldehyde
(1.3 g, 9.8
_,. _30__ _mmol)_was-added-dr~opwise-After_the_addition_was_complete,-stirring-
was-e.ontinued_at_
-70 C for another 15 minutes. Then 1.6M n-BuLi in hexanes (10 mL, 16 mmol)
was
added dropwise and stirring continued at -45 C for 4 h. The solution was
cooled to
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-70 C and then a solution of iodine (3.0 g, 12 mmol) and anhydrous THF (25
mL) was
added dropwise. When the addition was complete, stirring was continued at -70
C for 30
minutes and then at RT for 3 h. The crude product was talcen up in ether (40
mL) and
washed successively with saturated ammonium chloride (2 x 40 mL) and 5% sodium
5 thiosulfate (2 x 20 mL). The organic phase was dried, filtered and
concentrated.
Purification by column chromatography with EtOAc-heptanes (1:1) as eluent gave
0.41 g
(15% yield) of 4-iodo-6-methoxynicotinaldehyde.
LC-MS rnlz 264 (M+1);
1H NMR (CDC13) 8 9.95 (s, 1H), 8.53 (s, 1H), 7.32 (s, 1H) and 3.98 (s, 3H)
ppm.
4-lodo-6-methoxynicotinaldehyde (0.41 g, 1.6 nunol), trimethylsilylacetylene
(0.35 mL,
2.8 mmol), PdCl2(PPh3)Z (catalytic amount), CuI (catalytic amount), TEA (2 mL)
and THF
(10 mL) were stirred at 60 C for 2 h. The volatiles were evaporated and the
residue was
taken up in water and extracted with ether. The organic phase was dried,
filtered and
concentrated. Purification by column chromatography with EtOAc-heptanes (1:3)
as eluent
gave 0.25 g (68% yield) of 6-methoxy-4-
[(trimethylsilyl)ethynyl]nicotinaldehyde.
LC-MS m/z 234 (M+1);
H NMR (CDC13) 8 10.4 (s, 1H), 8.73 (s, 1H), 6.84 (s, 1H), 4.03 (s, 3H) and
0.30 (s, 9H)
ppm.
6-Methoxy-4-[(trimethylsilyl)ethynyl]nicotinaldehyde (0.25 g, 1.1 mmol) and 7M
ammonia in MeOH (5 mL) were stirred in a sealed vial at 80 C overnight. The
solution
was concentrated, taken up in saturated sodium carbonate and extracted with
ether. The
organic phase was dried, filtered and concentrated to give 0.20 g of the title
product.
GC-MS nalz 160 (M);
iH NMR (CDC13) b 9.41 (s, 1H), 9.27 (s, 1H), 8.47 (d, 1H), 7.64 (d, 1H), 7.03
(s, 1H) and
4:12 (s, 3H) ppm.
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Pharmacoloaical Examnle
Isolated Enzyme AssEs
MMP12
Recombinant human MMP12 catalytic domain may be expressed and purified as
described
by Parkar A.A. et al, (2000), Protein Expression and Purification, 20 152. The
purified
enzyme can be used to monitor inhibitors of activity as follows: MMP12 (50
ng/ml final
concentration) is incubated for 60 minutes at room temperature with the
synthetic substrate
Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 (10 M) in assay buffer (0.1M "Tris-HCl"
(trade marlc) buffer, pH 7.3 containing 0.1M NaCl, 20mM CaClz, 0.020 mM ZnCI
and
0.05% (w/v) "Brij 35" (trade mark) detergent) in the presence (10
concentrations) or
absence of inhibitors. Activity is determined by measuring the fluorescence at
kex 320 nm
and kem 405 nm. Percent inhibition is calculated as follows:
% Inhibition is equal to the [Fluorescenceplus inhibitor -
Fluorescencebackgf=ound] divided by
the [Fluorescenceninus inhibitor - Fluorescencebackgroundl=
MMP8
Purified pro-MMP8 is purchased from Calbiochem. The enzyme (at 10 g/ml) is
activated
by p-amino-phenyl==mercuric acetate (APMA) at 1 mM for 2.5 h, 35 C. The
activated
enzyme can be used to monitor inhibitors of activity as follows: MMP8 (200
ng/ml final
concentration) is incubated for 90 minutes at 35 C (80% H20) with the
synthetic substrate
Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 (12.5 M) in assay buffer (0.1M "Tris-HCl"
(trade marlc) buffer, pH 7.5 containing 0.1M NaCI, 30mM CaC12, 0.040 mM ZnCI
and
0.05% (w/v) "Brij 35" (trade mark) detergent) in the presence (10
concentrations) or
absence of inhibitors. Activity is determined by measuring the fluorescence at
kex 320 nm
and kem 405 nm. Percent inhibition is calculated as follows:
% Inhibition is equal to the [Fluorescenceplus inhibitor -
Fluorescencebackgf=ound] divided by
the [Fluorescenceminus inhibitor- Fluorescencebackgroundl=
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MMP9
Recombinant human MMP9 catalytic domain was expressed and then purified by Zn
chelate column chromatography followed by hydroxamate affinity column
chromatography. The enzyme can be used to monitor inhibitors of activity as
follows:
MMP9 (5 ng/inl final concentration) is incubated for 30 minutes at RT with the
synthetic
substrate Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 (5 M) in assay buffer (0.1M
"Tris-
HCI" (trade marlc) buffer, pH 7.3 containing 0.1M NaCl, 20mM CaCh, 0.020 mM
ZnCI
and 0.05% (w/v) "Brij 35" (trade marlc) detergent) in the presence (10
concentrations) or
absence of inhibitors. Activity is determined by measuring the fluorescence at
kex 320 nm.
and kem 405 mn. Percent inhibition is calculated as follows:
% Inhibition is equal to the [Fluorescenceplus inhibitor -
Fluorescencebackground] divided by
the [Fluorescence,,,inus inhibitoi - Fluorescencebackgroundl=
MMP14
Recombinant human MMP14 catalytic domain may be expressed and purified as
described
by Parkar A.A. et al, (2000), Protein Expression and Purification, 20 152. The
purified
enzyme can be used to monitor inhibitors of activity as follows: M1VIP14 (10
ng/ml final
concentration) is incubated for 60 minutes at room temperature with the
synthetic substrate
Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 (10 M) in assay buffer (0.1M "Tris-HCl"
(trade mark) buffer, pH 7.5 containing 0.1M NaCl, 20mM CaC12, 0.020 mM ZnCl
and
0.05% (w/v) "Brij 35" (trade mark) detergent) in the presence (5
concentrations) or
absence of inhibitors. Activity is determined by measuring the fluorescence at
kex 320 nm
and kem 405 nm. Percent inhibition is calculated as follows: % Inhibition is
equal to the
[Fluorescenceplus inlzibitor - Fluorescencebackground] divided by the
[FluorescenceõZinus
inhibitor - Fluorescencebackground]=
A protocol for testing against other matrix metalloproteinases, including
MMP9, using
expressed and purified pro MMP is described, for instance, by C. Graham Knight
et al.
z
(1992) FEBS Lett., 296(3), 263-266.
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MMP19
Recombinant human MMP19 catalytic domain may be expressed and purified as
described
by Parkar A.A. et al, (2000), Protein Expression and Purification, 20:152. The
purified
enzyme can be used to monitor inhibitors of activity as follows: MMP 19 (40
ng/n-A final
concentration) is incubated for 120 minutes at 35 C with the synthetic
substrate Mca-Pro-
Leu-Ala-Nva-Dpa-Ala-Arg-NH2 (5 M) in assay buffer (0.1M "Tris-HCl" (trade
inarlc)
buffer, pH 7.3 containing 0.1M NaCI, 20mM CaC12, 0.020 mM ZnCI and 0.05% (w/v)
"Brij 35" (trade mark) detergent) in the presence (5 concentrations) or
absence of
inhibitors. Activity is determined by measuring the fluorescence at kex 320 nm
and kem
405 nm. Percent inhibition is calculated as follows: % lnhibition is equal to
the
[Fluorescenceplus inhibitor= - Fluorescencebackground] divided by the
[Fluorescencenzinus
inhibitor- Fluorescencebackgrourzdl-
1s The following table shows data for a representative selection of the
compounds of the
present invention.
Table
hMMP12 hMMP9 hMMP14
Compound TCso (01) TCso (nM) ICso (nM)
Example 1 10.4 29.3 > 10000
Examp] e 2 1.4 3.5 415
Example 5 7 8.3 1990
